Isolation of Regenerating Hepatocytes after Partial Hepatectomy in Mice

The overall goal of this protocol is to present an optimized method for hepatocyte isolation in mice after partial hepatectomy without the need for density gradient centrifugation. We usually see a large percentage of lipid-laden hepatocytes during early regeneration. Those steatotic hepatocytes usually lost upon density gradient centrifugation due to the low density are retained with this protocol.

To prepare the perfusion equipment, connect a 26 gauge IUI cannula to the outlet end of the tubing using a Luer lock connector. Then flush the tubing and insert the inlet end of the tube into the pre-warmed perfusion buffer tube in the water bath. Prime it with warm perfusion buffer at a pump speed of three milliliters per minute.